Slieker W A, van der Loo J C, de Rijk-de Bruijn M F, Godfrey D I, Leenen P J, van Ewijk W
Department of Immunology, Erasmus University, Rotterdam, The Netherlands.
Int Immunol. 1993 Sep;5(9):1099-107. doi: 10.1093/intimm/5.9.1099.
In the accompanying paper we showed that six distinct subsets of bone marrow (BM) cells can be identified using the mAb ER-MP12 and ER-MP20 in two-colour immunofluorescence analysis. Upon intrathymic transfer into sublethally irradiated mice thymus-repopulating ability was restricted to ER-MP20- BM cells expressing either high or intermediate levels of the ER-MP12 antigen (1-2% and approximately 30% of BM nucleated cells respectively). The highest frequency of thymus-repopulating cells was found in the minor subset of ER-MP12(+)+20- BM cells. In the present study we demonstrate that upon intravenous transfer, thymus-homing and -repopulating BM cells are exclusively confined to the ER-MP12(+)+20- and ER-MP12+20- subpopulations, the highest frequency being detected among ER-MP12(+)+20- BM cells. Analysis of the peripheral blood leucocytes of reconstituted mice showed that not only prothymocytes but also progenitor cells of the B cell lineage as well as the myeloid lineage were present within both subsets. Three-colour flow cytometric analysis revealed that ER-MP12(+)+20- BM cells in particular were phenotypically heterogeneous with respect to the expression of the cell surface markers Thy-1, Sca-1, CD44, B220 and c-kit. Taken together our data demonstrate that ER-MP12 positively identifies BM cells with the ability to home to and repopulate the thymus. The phenotypic heterogeneity displayed by the ER-MP12(+)+20- BM subset, containing the highest frequency of thymus-homing and -repopulating cells, provides a basis for further separation of prothymocyte activity from other haematopoietic activities in the BM of the mouse.
在随附论文中我们表明,在双色免疫荧光分析中使用单克隆抗体ER-MP12和ER-MP20可识别出骨髓(BM)细胞的六个不同亚群。将其胸腺内移植到亚致死剂量照射的小鼠体内后,胸腺重建能力仅限于表达高水平或中等水平ER-MP12抗原的ER-MP20-BM细胞(分别占BM有核细胞的1-2%和约30%)。胸腺重建细胞的最高频率出现在ER-MP12(+)+20-BM细胞的较小亚群中。在本研究中我们证明,静脉内移植后,归巢至胸腺并重建胸腺的BM细胞仅局限于ER-MP12(+)+20-和ER-MP12+20-亚群,其中ER-MP12(+)+20-BM细胞中的频率最高。对重建小鼠外周血白细胞的分析表明,两个亚群中不仅存在前胸腺细胞,还存在B细胞谱系以及髓系谱系的祖细胞。三色流式细胞术分析显示,特别是ER-MP12(+)+20-BM细胞在细胞表面标志物Thy-1、Sca-1、CD44、B220和c-kit的表达方面具有表型异质性。综合我们的数据表明,ER-MP12可阳性识别具有归巢至胸腺并重建胸腺能力的BM细胞。含有最高频率胸腺归巢和重建细胞的ER-MP12(+)+20-BM亚群所显示的表型异质性,为从小鼠BM中的其他造血活性中进一步分离前胸腺细胞活性提供了基础。